Fig 3.

BoNT/D intoxication has a differential effect on HCR/A and HCR/T entry into neurons. BoNT/D (1 nM) was incubated with rat cortical neurons for 18 h. After being washed, neurons were incubated with 40 nM HA-HCR/T, HA-HCR/A, or Oyster 650 antisynaptotagmin antibody (anti-Syt, 1:400 dilution) for 5 min in low K buffer or high K buffer and then processed for IF. Representative images of indicated proteins after the 5-min incubation in high K buffer are shown. Relative intensities were normalized to the level for Syt; the high K signal was set to 1 within each data set. ns, not significant (P > 0.05); **, P < 0.01; ***, P < 0.001. Error bars indicate standard deviations.